1. Field of the Invention
The invention relates to mammalian kinase ligands, substrates and modulators. In particular, the invention relates to polypeptides, polypeptide compositions and polynucleotides that encode polypeptides that are ligands, substrates, and/or modulators of AKT. The invention also relates to polyligands that are homopolyligands or heteropolyligands that modulate AKT activity.
This application has subject matter related to application Ser. Nos. 10/724,532 (US 2004/0203027), 10/682,764 (US2004/0185556, PCT/US2004/013517, WO2005/040336), 11/233,246, and US20040572011P (WO2005116231). Each of these applications is hereby incorporated by reference.
2. Background of the Invention
Kinases are enzymes that catalyze the addition of phosphate to a molecule. The addition of phosphate by a kinase is called phosphorylation. When the kinase substrate is a protein molecule, the amino acids commonly phosphorylated are serine, threonine and tyrosine. Phosphatases are enzymes that remove phosphate from a molecule. The removal of phosphate is called dephosphorylation. Kinases and phosphatases often represent competing forces within a cell to transmit, attenuate, or otherwise modulate cellular signals and cellular control mechanisms. Kinases and phosphatases have both overlapping and unique natural substrates. Cellular signals and control mechanisms, as regulated by kinases, phosphatases, and their natural substrates are a target of research tool design and drug design.
Mammalian Protein Kinase B is also known as AKT. The enzymatic activity, activation and autoregulation of AKT have been studied. Several cellular substrates of AKT have been identified. A pharmacological agent that inhibits AKT activity has been disclosed in the literature by Martelli et al. (Leukemia (2003) 17:1794-1805). Natural and synthetic polypeptides have been studied to examine AKT substrate specificity. While polypeptides and variants thereof have been studied as individual AKT substrates or ligands, mixed ligands linked together as polyligands that modulate AKT activity have not been demonstrated before this invention. An aspect of the invention is to provide novel, modular, inhibitors of AKT activity by modifying one or more natural substrates either by truncation or by amino acid substitution. A further aspect of the invention is the subcellular localization of an AKT inhibitor, ligand, or polyligand by linking to a subcellular localization signal.
Design and synthesis of polypeptide ligands that modulate calcium/calmodulin-dependent protein kinase and that localize to the cardiac sarco(endo)plasmic reticulum was performed by Ji et al. (J Biol Chem (2003) 278:25063-71). Ji et al. accomplished this by generating expression constructs that localized calcium/calmodulin-dependent protein kinase inhibitory polypeptide ligands to the sarcoplasmic reticulum by fusing a sarcoplasmic reticulum localization signal derived from phospholamban to a polypeptide ligand. See also US 2004/0203027.
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